The analysis of glycoprotein structure and function, known as glycomics, has become a new and important area of research. Glycosylation is the most common post-translational modification of cell surface and extracellular matrix proteins. Glycoproteins play an important role in cell-adhesion and immune response. Changes in abundance and glycan profiles have been correlated with progression of diseases, such as cancer and rheumatoid arthritis. In addition, the analysis of glycan profiles is critical in the bio-therapeutic industry. For example, antibody bio-therapeutics contain glycosylated amino acids that assist in maintaining drug activity and preventing drug rejection by the immune system. Therefore, companies that make these bio-therapeutics have to monitor and verify their glycan profiles.
Glycan analysis is usually performed by capillary electrophoresis or mass spectrometry methods. In either case, the glycans must be removed from the glycoproteins, separated and analyzed. The process of removing glycans from glycoproteins traditionally includes an in-solution enzymatic reaction with PNGase F that requires a 24-hour incubation time. After the enzyme reaction, protein precipitation is needed to separate the glycans from the proteins for analysis of the glycans. Finally, the free glycans are analyzed with mass spectrometry or capillary electrophoresis. These steps are time-consuming and cumbersome, and the manual operations are error-prone. Furthermore, a relatively large quantity of starting materials is required to generate high quality data. Therefore, a better way of performing glycan analysis is desirable.